With advance of the molecular biology, various technologies are developed in relation with genes, and with those technologies, it is possible to separate and identify diseased genes, in much more numbers thereof. As a result, in the field of medical treatment, the molecular biological technologies are introduced into diagnoses or checking methods, thereby enabling the diagnosis, which was impossible in the conventional art, and also achieving great reduction in the number of days necessary for the checks.
Such the advance is mainly owned to an amplification method of a nucleic acid sequence, in particular, practicing of PCR (Polymerase Chain Reaction) technology or method. With this PCR method, it is possible to amplify the nucleic acid in a solution, specifically upon a sequence thereof. For this reason, it is possible to prove the existence of a virus living in blood serum of an extremely small quantity or amount, indirectly, by amplifying the nucleic acid sequences as the genes of the virus through the PCR method, for detection thereof. However, in a case where this PCR method is applied in daily clinical diagnoses in a clinical field, there still remain several problems to be dissolved. Among of them, a pre-treatment is very important, in particular, for estimation by using the PCR method, i.e., the capacity of refining of the nucleic acid sequence, and for this reason, there are proposed several technologies or methods in relation to the refining of the nucleic acid sequence.
For a first technology relating to the conventional art, a method is already disclosed, as described in Japanese Patent Laying-Open No. Hei 2-289596 (1990), for example, wherein silica particles, which can combine with the nucleic acid sequence under existence of chaotropic materials, are used as a solid-phase for use of combining or coupling with nucleic acid, for separating the solid-phase coupling with the nucleic acid sequence from a liquid, and thereafter complex of the solid-phase and the nucleic acid sequence (solid-phase/nucleic acid) is rinsed, thereby eluting the nucleic acid sequence from the complex, depending upon necessity thereof.
Also, for a second technology relating to the conventional art, a method is already disclosed, for example, in Japanese Patent Laying-Open No. Hei 8-320274 (1996), wherein a single nucleic acid sequence is extracted, collected, and/or separated, with using a large number of containers for a single sample, a large number of detachable tips for separate injection, filters, and particles of magnetic substance or material.
Further, for a third technology relating to the conventional art, a method is already disclosed, for example, in Japanese Patent Laying-Open No. 2000-514928 (2000) of PCT application, wherein blood plasma is separated from blood (i.e., whole blood) and further is mixed with an extremely small quantity of liquid, as well as detection of antibiotics, by using a microscopic channel, a chamber, a capillary, a deposable valve, etc.
However, the following problems can be listed, in the case where the methods or technologies disclosed in the conventional arts mentioned above are applied, actually.
First of all, with the method described in the Japanese Patent Laying-Open No. Hei 2-289596 (1990), mentioned as the first technology relating to the conventional art in the above, since the nucleic acid sequence can not be collected within the same device, it is difficult in automation thereof. Further, it is also difficult to increase the frequency of contact between the nucleic acid and the silica particles in a short time. In particular, in the case where the nucleic acid is low in the concentration, such as 102 copy/ml, for example, as a target contained within a sample, it is very difficult to increase the contact frequency in a short time.
Also, with the method described in the Japanese Patent Laying-Open No. Hei 8-320274 (1996), mentioned as the second technology relating to the conventional art in the above, processes are complex for extracting, collecting, and/or separating of the nucleic acid sequences, therefore the automation is difficult. Further, with this conventional art, since those processes cannot be treated within the same device, the problem of contamination thereof may be worried about.
On the other hand, in the second conventional art, it is disclosed that a pipette, to which the tip is connected, is linked with a cylinder (i.e., a syringe pump), so as to control the suction/discharge amount by means of a servomotor or a pulse-motor, severely. For this reason, for maintaining sufficient suction/discharge of the sample, it is impossible to increase the density of the silica-membrane filter. On the other hand, if applying a low density silica-membrane filter having a superior penetration of liquid, as a matter of course, the probability of collecting the nucleic acid becomes small. In particular, if the nucleic acid is low in the concentration, such as around 102 copy/ml, for example, as mentioned above, being the target contained within the sample, the contact frequency between the silica-membrane filter and the nucleic acids is reduced further, when the sample passes through that silica-membrane filter.
Also, in the Japanese Patent Laying-Open No. 2000-514928 (2000) of PCT application, as the third conventional art, there is disclosed the method for separating blood plasma from a very small amount of blood, such as about 150 μL, for example. However, in this publication, nothing is disclosed therein, about a technology nor a method for refining the nucleic acid.
Therefore, an object according to the present invention is to provide an automatic refining apparatus and a method thereof, which can be easily automated, wherein the contact frequency between the nucleic acids as the target within a sample and the solid-phase, thereby increasing trapping rate or ratio of the nucleic acids, even under the condition of the low concentration, such as around 102 copy/ml, for example, and further to provide a structure for refining nucleic acids for building up thereof, and to provide a gene analyzing apparatus and/or a refining apparatus for chemical materials with use thereof.